NOTES
May, 1963
337
TABLEI1 Acuw::, LETH.4L TOXICITY O F PIPERAZIXE-CoXT.4INING
C O M P O T ~ N D SIS hfICl7 AND
RATS
Total no. dead
--Compound a n d Vehicle
No. tested
Dose (mg./kg.)
24 hr.
33,000 30,250 27,500 18,857 13,469 9,620 3,204 3,360 2,240 1,120 8,000 6,000 4,000 3,000 12,000 8,000 6,000 4,000
19/30 3/10 0/l0 12/15 5/10 1/20 0/10 7/10 3/10 0/10 9/10 9/10 1/10 0/10 10/10 7/20 1/10 0/10
48 hr.
-LDso As compound
(mg,/kg.)a------As piperazine
Mouse DGP (I), 10%gum acaciab
1,4-Di(o-xylosyl)-piperazine (11),distilled H20
Anhydrous piperazine HC1 H20 ( p H 7.0)
+
Piperazine Hexahydrate HC1 -t H*O (pH 7.0) Piperazine citrate HC1 H20 (PH 6.0)
+
19/30 5/10 0/10 12/15 5/10 1/20 0/10 7/10 3/10 0/10 9/10 10/10 2/10 0/10 10/10 8/20 2/10 0/10
32,000 (33,280-30,866)
6,720 (6,989-6,482)
14,388 (16,953-12,384)
3,525 (4,153-3,034)
2,730 (3,331-2,238)
2,730 3,331-2,238)
5,300 (6,042-4,648)
2,385 (2,719-2,092)
8,500 (9,860-7,328)
3,400 (3,822-2,931)
Rat 8/10 30,000 33,000 7/10 6,300 3/10 3/10 (33,900-26,544) 27,500 (7,119-4,585) 1/10 0/10 22,000 10/10 10/10 11,200 15,000 4,502 Piperazine citrate 6/10 5/10 (12,678-9,825) 12,000 HC1 H20 ( p H 6.0) (5,097-3,950) 3/10 2/10 9,000 0/10 0/10 6,000 Acute Ln50 values iri distilled water were 29,900 mg./kg. (22,800a Figures in parentheses represent 95% confidence limits. 34,700 ), as piperazine 6,279 mg./kg. (4788-7287).
DGP (I) 10% gum acacia
+
‘
form” thereof for it t o exert oxyuricidal activity. The potency of the freshly prepared mixture was related directly t o its piperazine content and was essentially the same as DGP a t corresponding concentrations of piperazine. Although adult pinworms are more sensitive to the action of piperazine than are immature worms,10 all three compounds evaluated (Table I) showed significant activity against the immature worms, but DGP and anhydrous piperazine tended to show slightly more effect. Since the mouse pinworm is reportedly more resistant t o oxyuricides than the human pinworm,11 results of chemotherapeutic tests of such agents in mice are suggestive of their clinical activity. Based on piperazine content, D G P has essentially the same oxyuricidal potency as anhydrous piperazine and piperazine titrate. This equivalent activity with equivalent levels of piperazine has proved true x i t h all of the many soluble and insoluble piperazine derivatives1*that we have tested, provided that the compounds are altered in the body so as t o release piperazine or an “active form” thereof. Results obtained with the other glycosyl derivatives suggest that their solubility and/or hydrolyzability are of significance in their oxyuricidal activities. Toxicology.-Acute toxicity values were determined in mice and rats for D G P and piperazine citrate and in mice only for 1,4-di(D-xylosSl)piperazine (11), anhydrous piperazine and piperazine hexahydrate. The toxicity of the several agents \vas calculated in terms of the piperazine content which was 217, for DGP; 40% for piperazine citrate; 45% for piperazine hexahydrate and 1 0 0 ~ for o anhydrous piperazine. A repeated dose study using D G P as the test material \vas conducted in 6 week old albino rats. One of 4 groups, each consisting of 6 males and 6 females, served as controls. The remaining three groups represented dose levels of 3000, 2000, and (10) H. W. Brown, K. F. Chan, a n d K. L. Hussey, A m . J . T r o p . .Wed. Hug., 3, 504 (1954). (11) H. W. B r o u n , K. F. C h a n , a n d B. D. Ferrell, E x p . Parasztol., 3, 45 (1954); H. W. B r o u n and K. F. Chan, A m . J . T r o p . .Wed. Hug., 4, 3 2 1 (1955). (12) T. A. Martin, P. S. Prickett, a n d R’. T. Sumerford, unpublished results.
1000 mg. of DGP/kg. n-hich are equivalent to doses of 630, 420 and 210 mg. of piperazine/kg., respectively. D G P suspended in 107, gum acacia was administered orally once each day, 7 days a week for 10 weeks. Both 71-eights and food consumption values were recorded each week. Complete hematologic studies were conducted after 6 weeks of drug administration. A4cute, lethal toxicity of D G P in mice and rats is summarized in Table 11. Additional values for reference agents also are given. In general, acute, lethal doses of the piperazine derivatives produced ataxia, diarrhea and depression. Some deaths occurred between 24 and 48 hr. after dosing, but do not significantly alter the acute LDb, values. Repeated oral administration of DGP had no deleterious effect on food consumption or xeight gain. Hemograms for the treated animals revealed no marked differences from those of the control animals. Survival was not altered and gross examination of necropsy material revealed no pathologic changes attributable t o the drug. I n order to show further that the low order of toxicity exhibited by DGP is an inherent characteristic of the compound, some ~ 3 r k was 3 done with a mechanical mixture of 1 mole of piperazine and 2 moles of glucose. The LDjo value of the freshly prepared mixture as determined in white mice was found to be 2646 mg./kg. (based on the piperazine content) which is in the general range characteristic of piperazine and its salts. DGP v a s acutely less toxic than 1,4-di(~-sylosyl)piperazine, anhydrous piperazine, piperazine hexahydrate and piperazine citrate in mice and piperazine citrate in rats. Repeated doses of the product equivalent to 630 mg./kg. of piperazine each day were well tolerated in rats.
Acknowledgments.-We are indebted to Drs. J. R. Corrigan and W. T. Sumerford. We also thank the people who gave their assistance in the development of DGP: Miss D. Dansby, Messrs. H. L. Murray, R. L. Stratman, M. E. Fuquay, C. W. Stott, H. C. Hawkins, H. G. Payne, W. M. Coates, R. J. Bequette and R. R. Covington.
